Uncoupling protein, UCP, present in the inner mitochondrial membrane of brown adipose tissue (BAT) contributes to adaptive thermogenesis. UCP functions as a proton pore and can dissipate the proton electrochemical gradient established by the respiratory chain during fuel oxidation, and thus generates heat without producing ATP. However, the brown adipose tissue thermogenesis is not likely to be a major mechanism in controlling energy expenditure for humans because adults have only residual amounts of the tissue. Two new members of the UCP family have been identified based on their high sequence homology to UCP in BAT and named UCP2 and UCP3. The original UCP was renamed UCP1. At the amino acid level, human UCP2 and UCP3 are 59% and 57% identical to UCP1, respectively. In contrast to UCP1, UCP2 is expressed in many tissues such as brown adipose tissue, white adipose tissue, muscle, spleen and macrophages. UCP3 is expressed preferentially in skeletal muscle in humans, and brown adipose tissue and skeletal muscle in rodents. Since their identification many functional studies, including transgenic animals and ectopic expression of UCP2 or UCP3 in yeast, showed uncoupling activity of UCP2 and UCP3. A number of studies have been done that show increased expression of UCP2 and UCP3 by fasting, high-fat diets and suckling of newborn mice. A common characteristic of these circumstances is an associated increase in plasma free fatty acid levels. This study aimed to investigate effects of fatty acids, peroxisome proliferator-activated receptors (PPARs) and other transcription factors on UCP2 and UCP3 gene expression and to explore the molecular mechanism of their regulation through analysis of the promoter of the UCP2 and UCP3 genes. The 3.1 kb and 3.2 kb 5�-flanking regions of the mouse UCP2 and UCP3 genes, respectively, were cloned and used to construct promoter reporter gene (firefly luciferase) plasmids. The cloned region of the UCP2 and UCP3 genes contained putative binding motifs for several transcription factors, including PPAR, myogenin, and MyoD. Luciferase assays of both constructs showed basal promoter activity with 20~190-fold induction for the UCP2 promoter and 1.3~23-fold induction for the UCP3 promoter in several transfected cell lines, including 3T3-L1, C2C12, L6, COS7 and HepG2. Oleic acid (0.3 mM) up-regulated endogenous UCP2 mRNA by 2.3-fold in 3T3-L1 preadipocytes but not in C2C12 myotubes, and UCP3 mRNA by 2.5-fold in C2C12 myotubes. Responsiveness of the cloned promoter to oleic acid reflected the tissue-specific responsiveness of their endogenous genes but with less fold induction, 1.4-fold for UCP2 promoter in 3T3-L1 preadipocytes and 1.5-fold for UCP3 promoter in C2C12 myotubes. Forced expression of PPAR isotypes (PPARα, PPAR[delta] and PPARγ) showed tissue and isotype-specific activation of the UCP2 promoter. UCP2 promoter activity was induced by 2-fold by PPARγ in 3T3-L1 and by 2.8-fold by PPAR[delta] in C2C12. Treatment of oleic acid (0.3 mM) brought about further induction of the UCP2 promoter activity only in 3T3-L1. In contrast, all three isotypes induced activation of the UCP3 promoter in 3T3-L1, C2C12 and HepG2 cells. Treatment with oleic acid (0.3 mM) or isotype-specific agonist (10 [mu]M) resulted in further increased activity of the UCP3 promoter in 3T3-L1 and HepG2 cells. In particular, rosiglitazone (10 [mu]M) induced a 41-fold increase in UCP3 promoter activity in PPARγ transfected HepG2 cells, and this induction returned to basal level by treatment with bisphenol A diglycidyl ether (BADGE) (50 [mu]M), an antagonist for PPARγ. In addition, UCP3 promoter activity increased up to 20-fold 4 days after induction of C2C12 myoblasts differentiation, whereas UCP2 promoter activity increased only up to 2-fold. Forced expression of myogenin and MyoD in C2C12 myoblasts to mimic differentiation, induced UCP3 promoter activity in an additive manner, consistent with UCP3 being regulated by muscle differentiation. In the present study, it has been shown that UCP2 and UCP3 genes are regulated differently by fatty acids. The tissue-type dependence in regulation of endogenous UCP2 and UCP3 paralleled the cell type-specific effect of oleic acid on the promoter-reporter constructs, suggesting that fatty acid effects are at the transcriptional level. UCP2 and UCP3 promoters showed differences in their response to PPARs. Mediation of the fatty acid effect through PPARs has been also demonstrated, but direct binding of PPARs and particular regulatory motifs on the cloned promoter region have not yet been investigated.

Dietary omega-3 long chain polyunsaturated fatty acids (n-3 LCPUFAs), eicosapentaenoic acid (EPA, 20:5n-3), docosapentaenoic acid (DPA, 22:5n-3) and docosahexaenoic acid (DHA, 22:6n-3), have beneficial health effects and hence increasing the consumption of these fatty acids is recommended by health authorities. The most common dietary source of EPA, DPA and DHA is seafood, but few Australians habitually consume fish and on average eat less than one meal of fish per week. Thus if Australians are to meet the dietary guidelines for n-3 fatty acid intake, there is a need to develop a source of n-3 rich foods that fit into a typical Australian diet. Feeding fish oils rich in n-3 LCPUFA to chickens has proven problematic due to alteration in organoleptic properties. The incorporation of vegetable oils rich in n-3 PUFA, alpha-linolenic acid (ALA, 18:3n-3) into the diet of chickens is potentially an alternative way to provide meat rich in n-3 LCPUFAs as ALA is the precursor of EPA and DHA. However, most vegetable oils also contain the n-6 (n-6) PUFA, linoleic acid (LA, 18:2n-6) which competes with ALA for the same enzymes in their metabolism to LCPUFA. This thesis addressed two crucial issues relating to the conversion of ALA into EPA, DPA and DHA of chicken tissues. The objectives of the first experiment were to examine the effects of increasing the ALA content of diets on the conversion of ALA into EPA, DPA and DHA by measuring their accumulation in chicken meat (breast and thigh) and to determine if there was an optimum level of ALA (at a fixed level of LA) in this process. The ratio of LA to ALA of the diets ranged from 10.5:1 to 0.6:1. The findings in this study demonstrated that there was no optimum level of dietary ALA and as indicated by the observation that EPA, DPA and DHA continued to increase in breast and thigh as the ratio of LA to ALA decreased in the diet. In general, DPA achieved higher levels than DHA. The experimental diets with the lowest LA to ALA ratio elevated the incorporation of EPA and DHA into breast and thigh meat to levels 5 and 4-fold, respectively relative to birds fed the highest LA to ALA ratio. In contrast, arachidonic acid (AA, 20:4n-6) in all groups reduced with decreasing LA to ALA ratio in the diets. The results indicated that the dietary treatments did not significantly change the growth performance of chickens. The objective of the second experiment was to assess the regulatory effect of dietary LA on the conversion of ALA into EPA, DPA and DHA. While in the first experiment the diets varied in the level of ALA but had a constant LA level, in this experiment the level of ALA in the diets was held constant and the level of LA was varied. The LA to ALA ratio of experimental diets ranged from 1.4:1 to 2.1:1. The results of this study indicated that the highest LA to ALA ratio (2.1:1) resulted in the lowest n-3 LCPUFAs, EPA, DPA and DHA in meat samples. For example, the total n-3 LCPUFA levels in the breast meat of birds fed with the lowest LA to ALA ratio was 16% higher than the n-3 LCPUFA in the breast of birds fed the highest LA to ALA ratio. This study indicated that the strongest influence on EPA, DPA and DHA accumulation in chicken tissues was the level of ALA in the diet. The experimental diets did not appear to affect the growth performance of chickens. In conclusion, increasing the ALA content of chicken diets may result in a meat source high in n-3 LCPUFAs that may reduce pressure on diminishing marine stocks as well as offering health benefits to Australians. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1369065 / Thesis (M.Ag.Sc.) - University of Adelaide, School of Agriculture, Food and Wine, 2009

Erratum final three leaves of thesis. / Includes bibliographical references (leaves 266-288). / xvi, 288, [3] leaves : ill. ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Demonstrates that fatty acids and antioxidants can regulate the low density lipoprotein (LDL) receptor at the level of gene transcription in cultured liver cells. EPA and linoleic acid (PUFAs) are specifically shown to downregulate the LDL receptor compared to saturated and monosaturated fatty acids in the presence or absence of cholesterol. The experiments lead to the discovery that antioxidants can upregulate the LDL receptor in the human HepG2 cells. / Thesis (Ph.D.)--University of Adelaide, Dept. of Animal Science, 1996

Bibliography: leaves 242-298. / xxxiii, 298 leaves : ill. (some col.) ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / This thesis investigates the effects of polyunsaturated fatty acids (PUFAs) on macrophage oxygen radical production. The role of fatty acid structure in the ability to stimulate the fMLP response is also examined. The mechanisms by which fatty acids induce their effects on mononuclear phagocytes are partially elucidated. The mechanisms of the biological effects of the PUFAs in terms of intracellular signalling pathway are partly defined. / Thesis (Ph.D.)--University of Adelaide, Dept. of Paediatrics, 1997

Bibliography: leaves 242-298. / xxxiii, 298 leaves : ill. (some col.) ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / This thesis investigates the effects of polyunsaturated fatty acids (PUFAs) on macrophage oxygen radical production. The role of fatty acid structure in the ability to stimulate the fMLP response is also examined. The mechanisms by which fatty acids induce their effects on mononuclear phagocytes are partially elucidated. The mechanisms of the biological effects of the PUFAs in terms of intracellular signalling pathway are partly defined. / Thesis (Ph.D.)--University of Adelaide, Dept. of Paediatrics, 1997

Introduction: Gastric and colon cancer are major causes of mortality and morbidity worldwide. Gastric cancer is often detected at an advanced stage and current chemotherapeutics are only modestly effective against this neoplasm. Novel chemotherapeutics, chemopreventive agents and treatment strategies are required to prevent and treat gastric cancer. The ideal method to eliminate cancer cells may be the induction of apoptosis, further preventing cell proliferation and tumour growth. Recently, short-chain fatty acids (SCFAs) butyrate and propionate have been investigated as potential chemotherapeutic agents, particularly in colon cancer. Butyrate is reported to induce apoptosis in colon cancer cells and is demonstrated to modulate intracellular redox state by altering the levels of an antioxidant, glutathione (GSH). GSH availability is controlled by the oxidative pentose pathway (OPP). Very few studies have investigated the effects of butyrate on cell types other than colon cancer cells, and even less is known regarding the effects of propionate. This thesis investigated the potential for SCFAs to induce apoptosis in a gastric cancer cell line, Kato III, compared to the colon cancer cell line, Caco-2. Cell cycle regulation, OPP activity, GSH availability and glucose metabolism were also assessed. Methods: Initial studies developed a new technique to measure 1-13C-D-glucose metabolism. Following this, Kato III and Caco-2 colon carcinoma cells were treated with butyrate or propionate (1mM, 5mM or 10mM) or a 5mM combination of both SCFAs. The induction of apoptosis and cell cycle alterations by these SCFAs were assessed using flow cytometry. OPP activity and GSH availability were assessed in both cell lines using colorimetric techniques. Butyrate metabolism was assessed using 13C-butyrate. Results: Butyrate and propionate significantly induced apoptosis and G2-M arrest in Kato III and Caco-2 cells, although to a significantly greater extent in the latter cell line. Moreover, butyrate induced apoptosis to a significantly greater extent than propionate, in both cell lines. SCFA treatment led to the significant up-regulation of OPP activity in both cancer cell lines while GSH availability was significantly reduced. Glucose metabolism was initially increased by all SCFA treatments, however, 72hr butyrate treatment led to its reduction. Importantly, glucose metabolism was measured using a new technique developed within this thesis. The rate of butyrate metabolism was demonstrated to correlate with the sensitivity of each cell line to this SCFA. Conclusions: This thesis provides evidence that SCFAs, particularly butyrate, induce apoptosis in gastric and colon cancer cells in vitro. The response of cancer cells to SCFAs appears complex, and involves multiple distinct mechanisms and pathways, including p53, Fas, changes to intracellular redox state and glucose metabolism. The capability of butyrate to induce apoptosis also appears to be directly related to the rate of its metabolism. Butyrate has the potential to be utilised as an adjunctive therapy for the treatment of gastric cancer and colon cancer. / Thesis (Ph.D.) -- School of Molecular and Biomedical Science, 2007

Diss. (sammanfattning) Göteborg : Göteborgs universitet, 2007. / Härtill 4 uppsatser.

Thesis (M.S.)--West Virginia University, 2007. / Title from document title page. Document formatted into pages; contains vii, 44 p. : ill. Includes abstract. Includes bibliographical references (p. 30-33).

Thesis (Ph. D.)--University of Canterbury, 2006. / Typescript (photocopy). "December 2006." Includes bibliographical references (leaves 111-127). Also available via the World Wide Web.

Diss. (sammanfattning) Göteborg : Univ. , 2007. / Härtill 3 uppsatser.